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ob/ob小鼠是糖尿病相关研究中使用最广泛的动物模型之一,近年来,市场需求呈上升趋势。与野生型小鼠相比,其瘦素蛋白基因105号密码子发生C>T点突变,导致不能产生正常的瘦素蛋白。该模型4周前外观表型与野生型无异,而ob/ob纯合子不育,因而在繁殖建系过程中,需要通过基因型鉴定纯合、杂合和野生三种基因型。该文建立了基于高分辨率溶解曲线分析(high resolution melting analysis,HRM)的基因型鉴定方法,基因型分型结果与常规PCR加酶切方法一致,也与DNA测序结果吻合;通过该方法分型后获得纯合子小鼠外观表型、血糖浓度参数符合预期。该方法较常规方法省时、高效、节省实验成本,可用于大规模ob小鼠繁殖中的基因型鉴定。
Ob / ob mice are one of the most widely used animal models in diabetes-related research. In recent years, the market demand is on the rise. Compared to wild-type mice, codon 105 of the leptin gene undergoes a C> T point mutation, resulting in failure to produce normal leptin. The phenotype of the model was similar to that of the wild type 4 weeks ago, whereas the ob / ob homozygote was sterile. Therefore, three genotypes of homozygous, heterozygous and wild type were identified through genotyping during breeding and establishment. In this paper, a genotyping method based on high resolution melting analysis (HRM) was established. The results of genotyping were consistent with those of conventional PCR and DNA sequencing. By this method, After homozygous mouse appearance phenotype obtained, blood glucose concentration parameters in line with expectations. Compared with the conventional method, the method saves time, high efficiency and saves experimental cost, and can be used for genotyping in large-scale ob mice.